VviWRKY24 promotes β-damascenone biosynthesis by targeting VviNCED1 to increase abscisic acid in grape berries

Abstract

Norisoprenoids, which are produced by the cleavage of various carotenoids, are a class of volatile aroma compounds that widely distributed in plants. In wine, they represent a significant source of floral and fruity aromas. β-Damascenone is the most abundant and important norisoprenoid constituent in grape berries (Vitis vinifera L.) and wines. However, the regulatory mechanism of β-damascenone biosynthesis remains poorly understood. The present study has identified a WRKY transcription factor, VviWRKY24, as a key regulator of β-damascenone accumulation in grape berries. The results of overexpression and gene silencing assays in grape leaves, berries, and calli demonstrated that VviWRKY24 altered the flow of norisoprenoid metabolism and influenced the composition ratio of norisoprenoids, particularly enhancing the levels of β-damascenone. The results of the RNA-seq, yeast one-hybrid, electrophoretic mobility shift, and dual-luciferase assays provided confirmation that VviWRKY24 promoted abscisic acid (ABA) biosynthesis by directly upregulating the expression of VviNCED1. The increase in ABA content resulted in further induction of the expression of carotenoid cleavage dioxygenase 4B (VviCCD4b) on β-damascenone metabolic pathway. These findings elucidate the upstream regulation of ABA and the promotion of ABA on the accumulation of β-damascenone in grapes. This study contributes to a novel understanding of the regulatory mechanisms of β-damascenone biosynthesis and provides a strategy for improving the aroma quality of grapes and wine.

Acknowledgements

We thank Prof. Benzhong Zhu, Prof. Hongliang Zhu, Prof. Daqi Fu (China Agricultural University) for donating pTRV1 and pTRV2 vectors. We also thank Prof. Lei Sun (Beijing Academy of Agriculture and Forestry Sciences) for providing grape berries and V. quinquangularis leaves. This research was funded by the National Natural Science Foundation of China (grant nos 32072513 to Pan Q.H., U20A2042 to Duan C.Q., and 32102314 to Meng N.)

Author contributions

Y.W. performed the main experiments, interpreted data, and wrote the manuscript. Y.-C.W. assisted in the Y1H and dual-LUC and contributed to data interpretation. N.M. and H.-M.Z. participated in the transgenic calli experiment and the measurement of norisoprenoids. X.-C.Y., N.-Y.X., and X.M. contributed to the measurement of carotenoids and ABA. C.-Q.D. contributed to the research conception and design. Q.-H.P. conducted a critical review of the work and its scientific content. All authors contributed to the manuscript and approved the submitted version.

Data availability statement

Raw data of RNA-seq were uploaded in the SRA database and the accession number is PRJNA833286. Other data used in this study are available from the corresponding authors upon reasonable request.

Conflicts of interest

The authors declare no competing interest.

Supplementary Data

Supplementary data is available at Horticulture Research online.

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